Hydrogen [4-[[4-(diethylamino)phenyl][4-[ethyl[(3-sulphonatobenzyl)amino]-o-tolyl]methylene]-3-methylcyclohexa-2,5-dien-1-ylidene](ethyl)(3-sulphonatobenzyl)ammonium, sodium salt

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Remarks:

HPLC-MS/MS

Details on sampling:

At 0 h and at 48 h

Vehicle:

no

Details on test solutions:

- the necessary amount of test item for preparing the stock solution was weighed on a weighing scoop and transferred to a volumetric flask
- test medium (see Appendix B) was added up to the bench mark and the solution was homogenised by shaking
- the solution was clear and blue
- Lower test solutions were prepared by dilution of the appropriate solution with test medium
- ~50 mL of the prepared solutions were transferred to each test vessel

Test organisms (species):

Daphnia magna

Details on test organisms:

- Daphnia magna Straus, Clone V, was used as the test organism
- animals are continuously bred in the laboratory and were originally purchased in a healthy condition from the Federal Environment Agency in Berlin/Germany
- Daphnia magna was bred as single culture (1 daphnid per 100 mL) in Elendt M4 medium
- pHvalue of the aerated water was within a range of 6.0 – 9.0
- dissolved oxygen was above 60 % saturation and the total hardness 140 - 250 mg/L (as CaCO3), corresponding to 7.8 - 14°dH
- animals were fed with single cell green algae (Desmodesmus subspicatus, formerly Scenedesmus subspicatus) at least three times a week
- daphnids were reared at a temperature of 20 ± 2 °C in a climatic chamber with 16 hours of illumination and 8 hours of darkness
- medium was changed three times per week
- pipette was used to separate the young daphnids from the adults
- freshly hatched daphnids less than 24 hours old were used for the test

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

13 °dH (232mg/L as CaCO3)

Test temperature:

19.4 - 20.5 °C

pH:

7.7 - 8.1

Dissolved oxygen:

8.9 - 9.4 mg/L

Nominal and measured concentrations:

nominal (based on active ingredient): 116, 52.7, 24.0, 10.9, 4.95 mg/L (100, 45.5, 20.7, 9.39 and 4.27 mg/L)
The measured content of test item was between 93 % and 105 % of nominal in the fresh samples and between 98 % and 106 % in the aged samples.

Details on test conditions:

Test Conditions
- Test procedure: static
- Duration: 48 hours
- Temperature: 19.4 – 20.5 °C
- Oxygen concentration: ≥ 8.9 mg/L
- pH-value: 7.74 – 8.10
- Exposure to light: 16 hours photoperiod /8 hours darkness daily
- Feeding: none
- Test vessels: four 100 mL glass beakers with a glass plate per concentration, each filled with ~50 mL, one additional replicate for pysico - chemical measurements without test organisms
- Loading: ~10 mL of test solution for each animal
- Aeration: none
- Number of animals: 20 per concentration (4 replicates of 5 animals each)
- nominal concentrations: 100, 45.5, 20.7, 9.39 and 4.27 mg pure substance/L.
- Temperature, pH-value and oxygen concentration of the test solutions measured after 0, 24 hours aged and 48 hours are reported. Hardness of
the test water was measured on the day of application.
- criteria of effect: After 24 h and 48 h the immobilised daphnids were counted. All daphnids not able to swim within 15 seconds after gentle agitation of the test vessel were considered to be immobilised. If present, behavioural changes of daphnids were recorded at 24 and 48 hours after starting the test.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Details on results:

According to the results of the test, the EC50 (48 h) was supposed to be > 116 mg/L (nominal). Taking the purity of the test item into account, the EC50 (48 h) was supposed to be > 100 mg/L (nominal). The measured content of test item was between 93 % and 105 % of nominal in the fresh samples and between 98 % and 106 % in the aged samples.
After 24 hours and 48 hours of exposure in the test item concentrations of 52.7 (45.5) and 116 (100) mg/L daphnids were blue. Additionally at the test item concentration of 24.0 (20.7) mg/L the daphnids were slightly blue after 48 hours

Results with reference substance (positive control):

The results indicate an EC50 (24 h) of the reference item potassium dichromate between 1.00 mg/L and 2.00 mg/L.

Reported statistics and error estimates:

The values for EC50 could not be determined statistically since the immobilisation was below 50 % in the test.

Validity criteria fulfilled:

yes

Remarks:

immobilisation in control ≤ 10 %, dissolved oxygen concentration at the end of test should be ≥ 3 mg/L in all test units

Conclusions:

After 48 h of exposure to Special Brilliant Blue FFR no toxic effect against daphnia was observed. Thus a 48h-EC50 > 100 mg/L was determined.

Executive summary:

A study was conducted in accordance with OECD Guideline for Testing of Chemicals No. 202 'Daphnia sp., Acute Immobilisation Test' (adopted April 13, 2004) to assess the acute toxicity of Special Brilliant Blue FFR to Daphnia magna STRAUS.

The Daphnia were exposed to a range of concentration 100, 45.5, 20.7, 9.39 and 4.27 mg/L of the active ingredient of Special Brilliant Blue FFR for a period of 48 h. 20 daphnids (4 replicates a 5 animals) were exposed per concentration.

The test was conducted as a static test with the test medium unchanged throughout the duration of the test. During the test a temperature range of 19.4 - 20.5 °C was maintained in the test vessels. The temperature, the pH and the oxygen values were measured at the beginning, at 24 h and at the end of the test.

Observations were made on the swimming ability and the immobilisation rate, respectively, after 24 and 48 hours of exposure. After 48 h no toxic effects against Daphnia were observed for Special Brilliant Blue FFR under exposure conditions, thus no statistical analysis was required to determine the EC50 > 100 mg/L. After 24 hours and 48 hours of exposure in the test item concentrations of 45.5 and 100 mg/L daphnids were blue. Additionally at the test item concentration of 20.7 mg/L the daphnids were slightly blue after 48 hours. The results are expressed in terms of nominal concentrations based on active ingredient.

Description of key information

A study was conducted in accordance with OECD Guideline for Testing of Chemicals No. 202 'Daphnia sp., Acute Immobilisation Test' (adopted April 13, 2004) to assess the acute toxicity of Special Brilliant Blue FFR to Daphnia magna STRAUS.

The Daphnia were exposed to a range of concentration 100, 45.5, 20.7, 9.39 and 4.27 mg/L of the active ingredient of Special Brilliant Blue FFR for a period of 48 h. 20 daphnids (4 replicates a 5 animals) were exposed per concentration.

The test was conducted as a static test with the test medium unchanged throughout the duration of the test. During the test a temperature range of 19.4 - 20.5 °C was maintained in the test vessels. The temperature, the pH and the oxygen values were measured at the beginning, at 24 h and at the end of the test.

Observations were made on the swimming ability and the immobilisation rate, respectively, after 24 and 48 hours of exposure. After 48 h no toxic effects against Daphnia were observed for Special Brilliant Blue FFR under exposure conditions, thus no statistical analysis was required to determine the EC50 > 100 mg/L. After 24 hours and 48 hours of exposure in the test item concentrations of 45.5 and 100 mg/L daphnids were blue. Additionally at the test item concentration of 20.7 mg/L the daphnids were slightly blue after 48 hours. The results are expressed in terms of nominal concentrations based on active ingredient.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

Should read EC50 > 100 mg/L